CN109254002A - The identification signal of biomarker is converted into the labeling method and application of visible Optical Chromatography - Google Patents
The identification signal of biomarker is converted into the labeling method and application of visible Optical Chromatography Download PDFInfo
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- CN109254002A CN109254002A CN201811340761.6A CN201811340761A CN109254002A CN 109254002 A CN109254002 A CN 109254002A CN 201811340761 A CN201811340761 A CN 201811340761A CN 109254002 A CN109254002 A CN 109254002A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
- G01N33/533—Production of labelled immunochemicals with fluorescent label
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
Abstract
The invention discloses labeling methods and application that the identification signal of a kind of pair of biomarker is converted into visible Optical Chromatography, there is the liposome of power mutagens color characteristic using diacetylene monomer and phospholipid molecule preparation, the liposome is modified in the antibody for capableing of specific recognition marker to be measured, the probe molecules such as agglutinin or peptide molecule surface, probe molecule is reacted with target marker to be measured, cause the liposome that power mutagens color occurs, obtain the identification signal by probe molecule to biomarker to be measured, it is converted into the labeling method of macroscopic visible Optical Chromatography, it is qualitatively judged according to the color change that solution occurs.The method of the present invention and marker are in the apparent color change of visible-range generation, and required ingredient is less, and consumed cost is lower, and reaction process is faster.
Description
Technical field
The invention belongs to biomedical, clinical laboratory medicine and rapid detection technical fields, are related to a kind of pair of biological marker
The identification signal of object is converted into the labeling method and application of visible Optical Chromatography.
Background technique
As aging society arrives, orthopaedic disease shows the extent of injury day of human body and protrudes, and examines related disease early stage
The requirement that disconnected, process is grasped is also higher and higher.It is clear pathologically to recognize skeleton metabolism and related disease, for the life of bone
Reason state (bon e formation and absorption) comprehensive assessment is to judge bone development, bone decaying and bone loss, to examine bone disease
The basis of disconnected analysis.Wherein type i collagen is one of most important ingredient in bone structure, in bone tissue in bone matrix metabolism, I type
Collagen is degraded, and the molecule fragment of generation is released in blood or urine, these small fragments can effectively react bon e formation
With absorption.The tag fragments of reflection bon e formation have I type procollagen carboxypropeptide (PICP), reflect the marker piece of bone resorption
Section has type i collagen pyridine to be crosslinked whole peptide (ICTP) and type i collagen c-terminal peptides (CTX) etc., while other collagen types are degraded
The urine pyridine ether (PYD) and deoxidation pyridine ether (DPD) of generation are also the important indicator of reflection bone resorption.Therefore blood is quickly detected
Clearly, the above-mentioned molecule fragment of type i collagen is the main method of bone disease analyzing and diagnosing and the basis of quick diagnosis in urine.
It is comprehensive based on immune analysis detection method in many detection methods for being directed to type i collagen and its molecule fragment
Close the important method of assessment.Immunolabelling technique (IA) refers to the object for marking certain to be easy to detect in known antigens or antibody
Matter, by being converted into other signals for the specific binding of antigen and antibody, presence or absence or the concentration for detecting certain molecules are big
It is small.Due to the marker wide variety that this technology uses, the preparation of many products is simple, low in cost, therefore it is more
There is great application value in the different application field of kind.And the signal that different markers is converted out is also different, according to
Immunolabelling technique can be divided by marker difference, radio-immunity labelling technique, enzyme linked immunological labelling technique, chemiluminescence immunoassay
Labelling technique, fluorescence immunoassay labelling technique and colloidal gold immunolabeling technology etc..The common problem of these labeling methods is not
Suitable for quickly detecting and semi-quantitative analysis, it is particularly unsuited for community hospital and the home tele-monitoring of aging society.It enumerates below
Existing labelling technique there is shortcoming:
(1) radio-immunity labelling technique: there are faint radioactive pollution, waste disposal difficulty is big；Radioactive isotope
The presence of half-life period increases experimental implementation difficulty；
(2) enzyme linked immunological labelling technique: enzyme marker is easy inactivation；Sensitivity is not high；Reaction process is relative complex；
(3) fluorescence immunoassay labelling technique: reaction process is rapid, but simultaneously vulnerable to light scattering and sample Intrinsic fluorescence
The influence of substance；
(4) electrochemiluminescent immunoassay labelling technique: needing additional solid phase carrier, and operating process is relative complex；
(5) colloidal gold immunolabeling technology: sensitivity is impacted, and technology wants high, and operating process is difficult.
In conclusion at present because label bring is unfavorable for basic hospital and family's fast diagnosis method, the hair
It is bright to be based on immunoassay, a kind of novel markings method is designed, to the quick diagnosis of type i collagen molecule fragment, bed in urine, serum
Preceding diagnosis (POCT) is particularly important to child, old man.Comparison various not isolabelings and analysis method, are based on using one kind
Power causes color-change technology, becomes the conversion regime of visible light colors signal, and be prepared into color spectrogram or color board, is suitable for family
Front yard give birth to children with endowment and it is other need it is simple, quickly and the place of science judgment.
Summary of the invention
It is an object of the invention to establish a kind of new immune labeled method, color-change technology is caused using power, by antibody antigen
Between specific binding affinity be converted into macroscopic optical signal.This visible pass optical signal characterized by color change, thus
Design a kind of novel immune analytical technology suitable for quickly detecting type i collagen.
The present invention is realized especially by following technical scheme:
A kind of immune labeled method suitable for field quick detection, comprising:
Using diacetylene structures alone and phospholipid molecule aggregation, it is visible light signal lipid that preparation, which has power mutagens color,
Body；
The liposome is modified with the antibody, agglutinin or peptide molecule for capableing of specific recognition determinand in lipid body surface
Face obtains biomarker；
Target determinand is reacted with biomarker, is qualitatively judged according to the color change that solution occurs.
The diacetylene structures alone is selected from bis- pentadecane diacetylenic acid of 10,12-, 23 carbon diacetylenic acid of 10,12-, 10,
25 diine -1- alcohol of 12-, 2,4- pentadecane diacetylenic acid, 19 carbon diacetylenic acid of 2,4-, 17 carbon diacetylenic acid of 2,4-, 10,12-
27 carbon diacetylenic acids, 10,12- pentacosane diacetylenic acid methyl esters, 29 carbon diacetylenic acid of 10,12-, 16 carbon diacetylenic acid of 5,7-
One of kind.
The liposome by the way that the one or more of diacetylene structures alone are dissolved in solvent, or by monomer with
It is dissolved after phospholipid molecule mixing, prepares the liposome of different sizes and form.
Preferably, the specific preparation process of the liposome is in a solvent by diacetylene structures alone and different second
Alkynes structures alone or phospholipid molecule mixing, are configured to solution and remove solvent, add buffer and keep concentration, by the dispersion
Liquid saves after ultrasonic vibration under certain condition.
Wherein, the buffer solution is selected from phosphate, citrate, carbonate, borate, barbiturates, three hydroxyl first
Any one in base aminomethane alkali or acetate buffer.
Preferably, in such a way that film filters, separate the different liposome of the size modes, leave size 30~
The liposome of smooth chondritic between 250nm.
Preferably, the method system that the liposome blows out film ultrasonic method using injection method or rotating thin film ultrasonic method or nitrogen
It is standby.
Preferably, for the aggregation of diacetylene structures alone assembling, the modification means are selected from insertion, absorption or even
Connection.Wherein the crosslinking agent of chemical coupling is planted selected from one of carbodiimide, glutaraldehyde, glutamine transaminage, Geniposide.
Preferably, the aggregation being mixed with for diacetylene structures alone and phospholipid molecule, the modification means are
Probe molecule (ligand) is embedded in phospholipid molecule and is obtained.
Using the method for the present invention by the antibody assembly of various concentration in surface of liposome, with obtained antibody/liposome life
Substance markers analyte detection identifies the object of various concentration, draws out color spectrogram or color board according to color gradient.
In another aspect of this invention, the present invention provides a kind of biomarker, the biomarker pass through by
Antibody, agglutinin or the peptide molecule for capableing of specific recognition determinand are assemblied in diacetylene structures alone and phospholipid molecule aggregation
The surface of body obtains.
In another aspect of this invention, the present invention provides a kind of for marking type i collagen and its relevant molecule segment
Biomarker, is mixed with aggregation by diacetylene structures alone and phospholipid molecule, and probe molecule is embedded in phosphatide point
It is obtained in son.
Wherein, the probe molecule is type i collagen monoclonal antibody, I type procollagen carboxypropeptide (PICP) Dan Ke
Grand antibody, type i collagen pyridine are crosslinked whole peptide (ICTP) monoclonal antibody, type i collagen c-terminal peptides (CTX) monoclonal antibody, urine
Pyridine ether (PYD) monoclonal antibody or deoxidation pyridine ether (DPD) monoclonal antibody.
The type i collagen relevant molecule segment is I type procollagen carboxypropeptide (PICP), the crosslinking of type i collagen pyridine
Whole peptide (ICTP), type i collagen c-terminal peptides (CTX), urine pyridine ether (PYD) or deoxidation pyridine ether (DPD) etc..
In another aspect of this invention, a kind of for quickly detecting the immune labeled of type i collagen and its relevant molecule segment
Method, by test serum or the above-mentioned biomarker of urine sample addition containing a certain amount of type i collagen or relevant molecule segment
In, constant-temperature incubation for a period of time, observes color change, compares color board, quickly obtains target concentration size.
In another aspect of this invention, above-mentioned biomarker makees analyte detection type i collagen and its relevant molecule segment is being received
Application in rice sensor.
In another aspect of this invention, a kind of novel immune labelling technique based on fluorescence signal conversion can also be provided.
The application method of the novel immune labelling technique based on fluorescence signal conversion includes: will be containing a certain amount of I type
Tropocollagen molecule or the test serum or urine sample of relevant molecule segment are added in above-mentioned biomarker tool, and one section of constant-temperature incubation
Time observes mixed liquor color change, and the color change is by tropocollagen molecule in sample or the shadow of small molecule segment concentration
It rings, liposome fluorescence signal changes, and carries out semi-quantitative analysis using Laser Scanning Confocal Microscope.
The invention has the benefit that
The present invention provides a kind of novel markings method for being different from traditional immunization labelling technique, for current traditional immunization mark
According to Immune discrimination technology, and power mutagens color is occurred by conjugated polymer in visible-range for defect existing for note technology
Mechanism etc., establish a kind of novel markings means for being different from traditional immunization labelling technique.The present invention has easy to operate, reaction
Rapidly, stability is high, signal feature easy to identify, be it is a kind of more efficient than traditional biological labeling method, be easier to popularize, cost
Less expensive novel markings technology.The present invention is using antibody, agglutinin or peptide molecule to the specific recognition of corresponding object
And the mechanism of power mutagens color occurs in visible-range for molecule aggregate, which, which is changed into, visually to identify
Optical signal, so as to carry out quantification and qualification to object.
The present invention has simple, quick, stable, high as a kind of novel markings technology for having efficient identification object
The characteristics of effect；Designed new bio detects identification facility in the present invention, and the signal after transformation occurs bright in visible-range
Aobvious color change, more conducively naked eyes identify；New biomarker technology designed by the present invention, compared to traditional Immune discrimination
Technology, required ingredient is less, and consumed cost is lower, and reaction process is faster.
Specific embodiment
Below in conjunction with specific embodiment of the present invention, technical solution of the present invention is clearly and completely described, is shown
So, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Based on the reality in the present invention
Example is applied, every other embodiment obtained by those of ordinary skill in the art without making creative efforts all belongs to
In the scope of protection of the invention.
Amphiphilic monomer containing diacetylene structure can be self-assembly of liposome structure, can occur under ultraviolet lighting
Polymerization reaction, so that intermolecular rigid segment of the formation containing conjugated double bond.The bilayer molecules membrane structure of surface of liposome is similar
Biomembrane, before polymerization the characteristics of having both stability and mobility simultaneously.And in forming process, because there are charges for film surface
Relationship so that film closure formed liposome simultaneously easily occur agglomeration.Since there are above two features, so that should
The liposome that amphiphilic monomer is self-assembly of has the mutually different structure of several shape sizes.In the liposome of above structure
In, in 30-250nm, the smooth spherical liposome of surface texture has outstanding power and causes discoloration size.Work as liposome
When surface is by external force, the intermolecular rigid segment containing conjugated double bond structures is orientated along Impact direction, thus
So that the cloud density of the conjugated double bond in entire membrane structure changes, migrate it to the absorption of visible light.?
Macroscopically it is presented as macroscopic color transition.Therefore, which, can be in visible-range when by external force
The interior rapid apparent discoloration of generation, has excellent signal changing function.When it is used for immune labeled field, can effectively improve
Traditional immunization labelling technique signal identification is difficult, the slow disadvantage of reaction process.Meanwhile liposome meeting when by environmental stimuli
There is the transformation of fluorescence signal, can be equally used for immune labeled field.
Further, since the characteristics of bilayer molecules membrane structure of surface of liposome has both stability and mobility simultaneously, when it
When as immune labeled field, solid phase carrier and signal converter can be played the role of simultaneously.Therefore, immune labeled skill is utilized
Art, will antibody, agglutinin or the peptide molecule of specific recognition target determinand change in conjunction with determinand, then by the signal
For the optical signal that can visually identify in visible-range, or the fluorescence signal easily detected by instrument, reach qualitative or semidefinite
Measure the purpose quickly detected.
In novel markings method designed by the present invention, required biomarker the preparation method is as follows:
First in a solvent by the dissolution of diacetylene structures alone, super using film by injection method or between 55~75 DEG C
Sound method prepares the liposome of three kinds of different sizes and form, is kept in dark place after crosslinking.
The different liposome of the size modes is separated by way of dialysing or filtering, and leaves size in 30~250nm
Between smooth chondritic liposome.
Using assembling means such as insertion, absorption or couplings, by the liposome be capable of specific recognition determinand antibody,
Agglutinin or peptide molecule are assembled, and are obtained a kind of novel markings tool for being different from traditional immunization labelling technique, are protected from light 4 DEG C
It saves.
Wherein diacetylene structures alone be bis- pentadecane diacetylenic acid (10,12-Pentacosadiynoicacid) of 10,12-,
23 carbon diacetylenic acid (10,12-Tricosadiynoicacid) of 10,12-, bis- pentadecane diine -1- alcohol (10,12- of 10,12-
Pentacosadiyn-1-ol), 2,4- pentadecane diacetylenic acid (2,4-pentadecadiynoic acid), 19 carbon two of 2,4-
Acetylenic acid (2,4-nonadecadiynoic acid), 17 carbon diacetylenic acid of 2,4- (2,4-heptadecadiynoic acid),
27 carbon diacetylenic acid of 10,12- (10,12-heptacosadiynoic acid), 10,12- pentacosane diacetylenic acid methyl esters
(Methyl-10,12-pentacosadiynoate), 29 carbon diacetylenic acid (10,12-nonacosadiynoic of 10,12-
Acid), any one in 16 carbon diacetylenic acid of 5,7- (5,7-hexadecadiynoic acid).
Solvent is solution phosphate, citrate, carbonate, borate, barbiturates, Tris alkali (three hydroxyls in liposome
Aminomethane alkali) or acetate buffer in any one.
Chemical coupling in assembly is that the coupling agent is carbodiimide, HOSu NHS, glutaraldehyde, glutamine
Any one in transaminase, Geniposide, reaction temperature is maintained between 4-37 DEG C.
The operational means of the novel markings method of target determinand is as follows for identification in the present invention:
A certain amount of tissue containing target determinand, serum or urine sample are added in above-mentioned biomarker, it is permanent
Temperature is incubated for a period of time, observation solution colour variation, which is influenced by target testing concentration size in sample,
Solution colour can change with fluorescence signal, carry out semidefinite using ultraviolet-visible spectrophotometer or Laser Scanning Confocal Microscope
Amount analysis.
Present invention uses the amphiphile, amphiphilic molecules with diacetylene structure to make it from group using the special construction of amphiphile, amphiphilic molecule
Dress forms liposome, using the special construction of polymerized liposome molecular backbone of the crosslinking after fixed, when extraneous stress acts on rouge
When on the molecular side chain of liposome surface, Cloud Distribution in conjugated double bond can be influenced on main chain, eventually leads to liposome hair
Raw macroscopic color change.Using this discoloration, by antibody coupling in surface of liposome, with testing molecule specificity
In conjunction with the external force of generation acts directly on polymer molecule side chain, so as to cause liposome color change.Pass through comparison front and back ratio
The content of the detectable determinand of the variation of coloration.In addition, liposome can equally generate fluorescence signal when by external forces
Transformation, can pass through comparison front and back fluorescence intensity detect determinand content.
Application of the invention is described further below with reference to specific embodiments of the present invention:
Embodiment 1
Bis- pentadecane diacetylenic acid of 10,12- (10,12-Pentacosadiynoic acid, PCDA) is dissolved in dehydrated alcohol
In, liposome is prepared using film ultrasound.It is PBS buffer solution (10mM) that solvent in liposome solutions, which is made, and liposome is dense
Degree is 1mM.It is kept in dark place at 4 DEG C overnight, UV crosslinking obtains blue polymer liposome.It will specific recognition β isomery CTX
The polyclonal antibody of segment is dissolved in the PBS buffer solution that concentration is 10mM, is mixed with liposome solutions, and carbodiimide is added
(EDC) with n-hydroxysuccinimide (NHS) (concentration ratio EDC:NHS:liposome=2:1:1), by probe conjugate in lipid
Body surface face is centrifuged off the antibody to dissociate in solution, is kept in dark place.
It is that sample of the 1mg/ml containing target detection thing is added by 6 μ l concentration, is incubated for 30 minutes at 25 DEG C, color is become by indigo plant
It is red.
Embodiment 2
29 carbon diacetylenic acid of 10,12- (10,12-nonacosadiynoic acid) is dissolved in chloroform and phosphatide point
Son mixing, prepares liposome using injection method.It is PBS buffer solution (10mM) that solvent in liposome solutions, which is made, and liposome is dense
Degree is 0.5mM.It is kept in dark place at 4 DEG C overnight, UV crosslinking obtains blue polymer liposome.It will specific recognition CTX piece
The polyclonal antibody of section is dissolved in the PBS buffer solution that concentration is 10mM, is mixed with liposome solutions, and mass fraction 25% is added
Glutaraldehyde, by probe conjugate in surface of liposome, be centrifuged off in solution dissociate antibody, be kept in dark place.
It is that sample of the 1mg/ml containing target detection thing is added by 4 μ l concentration, is incubated for 30 minutes at 25 DEG C, color is by leucismus
It is red.
Embodiment 3
10,12-, 23 carbon diacetylenic acid (10,12-Tricosadiynoicacid) is dissolved in chloroform, it is super using film
Sound method prepares liposome.It is PBS buffer solution (10mM), concentration of liposomes 0.5mM that solvent in liposome solutions, which is made,.4℃
Under be kept in dark place overnight, UV crosslinking obtains blue polymer liposome.Will specific recognition I-type collagen it is polyclonal
Antibody is dissolved in the Tris buffer that concentration is 10mM, is mixed with liposome solutions, and the glutaraldehyde of mass fraction 25% is added,
By probe conjugate in surface of liposome, it is centrifuged off the antibody to dissociate in solution, is kept in dark place.
It is that sample of the 0.5mg/ml containing target detection thing is added by 6 μ l concentration, is incubated for 30 minutes at 25 DEG C, color is become by ash
It is purple.
Embodiment 4
By 23 carbon diacetylenic acid (10,12-Tricosadiynoicacid) of 10,12- and 2,4- pentadecane diacetylenic acid (2,
4-pentadecadiynoic acid) it is dissolved in chloroform, liposome is prepared using film ultrasound.Liposome solutions are made
Middle solvent is buffer (10mM), concentration of liposomes 0.5mM.It is kept in dark place at 4 DEG C overnight, UV crosslinking obtains light blue polymerization
Liposome.Will specific recognition I-type collagen polyclonal antibody be dissolved in concentration be 10mM Tris buffer in, with
Liposome solutions mixing, the Geniposide that mass fraction 25% is added are centrifuged off in solution by probe conjugate in surface of liposome
Free antibody, is kept in dark place.
It is that sample of the 0.5mg/ml containing target detection thing is added by 6 μ l concentration, is incubated for 30 minutes at 25 DEG C, color is by light blue
Purpling.
Embodiment 5
By 10,12- pentacosane diacetylenic acid methyl esters (Methyl-10,12-pentacosadiynoate) and 2,4- 15
Carbon diacetylenic acid (2,4-pentadecadiynoic acid) and phospholipid molecule are miscible in chloroform, are prepared using film ultrasound
Liposome out.It is buffer (10mM), concentration of liposomes 0.3mM that solvent in liposome solutions, which is made,.It was kept in dark place at 4 DEG C
Night, UV crosslinking obtain colourless polymerized liposome.Will the polyclonal antibody of specific recognition I-type collagen be dissolved in concentration
To be mixed in the Tris buffer of 10mM with liposome solutions, the genipin solution of mass fraction 20% is added, by probe conjugate
In surface of liposome, it is centrifuged off the antibody to dissociate in solution, is kept in dark place.
It is that sample of the 0.5mg/ml containing target detection thing is added by 6 μ l concentration, is incubated for 30 minutes at 25 DEG C, color is by colourless
It reddens.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
Understand without departing from the principles and spirit of the present invention can to these examples carry out it is a variety of variation, modification, replacement and
Modification, the scope of the present invention is defined by the appended.
Claims (10)
1. the labeling method that the identification signal of a kind of pair of biomarker is converted into visible Optical Chromatography characterized by comprising
Using diacetylene monomer and phospholipid molecule, it is visible light signal liposome that preparation, which has power mutagens color,；
By the liposome modify or mark with antibody, agglutinin or the polypeptide etc. of being capable of specific recognition biomarker to be measured
Probe molecule surface, obtains biosensor；
Target biomarker to be measured and probe molecule specific reaction cause the liposome that power mutagens color occurs, according to solution
The color change of generation is qualitatively judged.
2. the label side that the identification signal of a kind of pair of biomarker according to claim 1 is converted into visible Optical Chromatography
Method, which is characterized in that the diacetylene structures alone is selected from 10,12-, bis- pentadecane diacetylenic acid, 10,12-, 23 carbon diine
Acid, 25 diine -1- alcohol of 10,12-, 2,4- pentadecane diacetylenic acid, 19 carbon diacetylenic acid of 2,4-, 17 carbon diacetylenic acid of 2,4-,
27 carbon diacetylenic acid of 10,12-, 10,12- pentacosane diacetylenic acid methyl esters, 29 carbon diacetylenic acid of 10,12-, 16 carbon of 5,7-
One of diacetylenic acid kind.
3. the label side that the identification signal of a kind of pair of biomarker according to claim 1 is converted into visible Optical Chromatography
Method, which is characterized in that the liposome, or will by the way that the one or more of diacetylene structures alone are dissolved in solvent
Monomer dissolves after mixing with phospholipid molecule, and liposome is prepared.
4. the label side that the identification signal of a kind of pair of biomarker according to claim 3 is converted into visible Optical Chromatography
Method, which is characterized in that for the aggregation of diacetylene structures alone assembling, the modification means are insertion, absorption or coupling.
5. the label side that the identification signal of a kind of pair of biomarker according to claim 3 is converted into visible Optical Chromatography
Method, which is characterized in that for the aggregation that diacetylene structures alone and phospholipid molecule are mixed with, the modification or label hand
Section obtains for probe molecule to be embedded in phospholipid molecule.
6. a kind of for marking the biomarker of type i collagen and its relevant molecule segment, which is characterized in that pass through diacetylene knot
Structure monomer and phospholipid molecule are mixed with aggregation, and probe molecule is embedded in phospholipid molecule and is obtained.
7. according to claim 6 for marking the biomarker of type i collagen and its relevant molecule segment, feature exists
In the probe molecule is type i collagen monoclonal antibody, I type procollagen carboxypropeptide monoclonal antibody, type i collagen pyrrole
Pyridine is crosslinked whole peptide monoclonal antibody, type i collagen c-terminal peptides monoclonal antibody, urine pyridine ether monoclonal antibody or deoxidation pyridine ether
Monoclonal antibody.
8. according to claim 6 for marking the biomarker of type i collagen and its relevant molecule segment, feature exists
In the type i collagen relevant molecule segment is I type procollagen carboxypropeptide, and type i collagen pyridine is crosslinked whole peptide, type i collagen
C-terminal peptides, urine pyridine ether or deoxidation pyridine ether.
9. a kind of for quickly detecting the labeling method of type i collagen and its relevant molecule segment, which is characterized in that will contain a certain amount of
Type i collagen or the test serum or urine sample of relevant molecule segment be added in biomarker as claimed in claim 6, it is permanent
Temperature is incubated for a period of time, observes color change, compares color board, quickly obtains target concentration size.
10. biomarker as claimed in claim 6 makees analyte detection type i collagen and its relevant molecule segment in nano-sensor
Application.
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| CN110208550A (en) * | 2019-07-03 | 2019-09-06 | 贵州省临床检验中心 | One kind marker relevant with risk of recurrence after Atrial fibrillation radiofrequency ablation is combined and its is applied |
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